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Testicular endocrine function was studied in twelve post pubertal patients with male pseudohermaphroditism and 46 XY chromosomal constitution. Patients were divided into three groups, four subjects who became feminized during puberty, five who became masculinized during puberty and three who were castrated before puberty. Serum dehydroepiandrosterone, progesterone, 17-hydroxyprogesterone, androstendione, testosterone, dihydrotestosterone, LH and FSH were determined by radioimmunoassay. Patients of the first group had the clinical characteristics of testicular feminization secondary to absence of the androgen receptor. One of the five patients of the second group had deficient testosterone secretion but no enzymatic defect could be demonstrated. One of the subjects castrated before puberty had a deficiency in 17,20-desmolase. Therefore, evidence of a failure of the fetal testes could be found in only two of the twelve patients studied.

A case of retrograde ejaculation not responding to medical treatment is described in which the problem of sterility was overcome by artificial insemination. The semen was collected from urine after coitus by previous alkalinization with 1.63 g NaHCO3 once daily. Pregnancy occurred at the 4th insemination during the 1st treatment cycle. This technique seems to be the most suitable for artificial insemination in cases of retrograde ejaculation.

We have used GIFT in 74 couples affected with unexplained sterility, male hypofertility, endometriosis, phymosis of the fimbria and adhesions impeding oocyte pick-up. We systematically desensitized the pituitary using a GnRH analog from the 20th day of the cycle preceding GIFT. All the patients were hyperstimulated with purified FSH and hMG (Metrodin and Pergonal-Serono) from the 3rd day of the cycle. When several follicles were obtained woth a diameter greater than 16 mm together with serum estradiol values of 300 pg/ml per follicle, 10,000 IU hCG were administered intramuscularly in a single dose (Profasi-Serono). A maximum of 5 oocytes per patient were transferred (2-3 per tube). Our results confirm the validity of the GIFT method, with an average success rate of 19% in all pathological conditions and a far greater success rate in couples with unexplained sterility than in those with male hypofertility (31% and 10% respectively). The Authors stress that the best results of GIFT were achieved when the ratio between E2 and follicles with a diameter greater than 16 mm was in excess of 500 pg/ml.

The authors administrated the Clomiphene, in different doses, to 8 patients among 27 observed. These patients were checked for a long time by basal body temperature and in some cases, measuring total urinary estrogens and pregnandioluria. The therapy was administrated in 38 cicles of the 163 cicles observed. This drug got evident results on flux duration, less significant results on the cicle length. The Clomiphene produced ovulatory cicles in 58 per cent of the cases, against 7 per cent of spontaneous ovulatory cicles of the same patients. The drug effect is better in the cases with normo or moderate hypo or hyper estrogenism, not so good in the cases with a remarcable hypoestrogenism. The therapy with Clomiphene did not produce any inconvenient. The results got were put in comparison with those of the other drug (Ciclophenile) and the superiority of Clomiphene was confirmed.

Ovarian resection was performed by means of celioscopy in 12 patients with polycystic ovaries (PCO), who had not responded to clomiphene + HCG therapy. This therapy had induced ovulation in only 14% of 56 therapeutic cycles. Ovarian resection through celioscopy induced ovulation in 45% of cycles. Clomiphene + HCG therapy after resection in patients with no spontaneous ovulation induced ovulation in 44% of cycles. Seven pregnancies followed in 5 women, equal to 41% of patients. Hormonal patterns (FSH, LH, E2, progesterone, dehydroepiandrosterone sulfate, testosterone) before and after resection showed no significant variation. The authors emphasize other advantages of this technique, such as fewer surgical injuries than laparotomy, relative harmlessness of the treatment, the possibility of repeating it and of performing subsequent laparotomy.

On different days following coitus, i.e., during the first stages of pregnancy, adult female rabbits were treated with antiblastokinin obtained from a chicken. A high prenatal mortality was observed and a reduction of the weight in the stillborn of the rabbits treated. The antiblastokinin produces evident biological effect about the 4th day following coitus, during the period of maximum production of blastokinin by the endometrium in vivo.

Purified urinary follicle-stimulating hormone was used to induce ovulation in 18 patients with polycystic ovarian syndrome. Each ampule contained 75 IU of follicle-stimulating hormone and less than 0.11 IU of luteinizing hormone. Initial doses were 150 to 225 IU/day, later increased to a maximum of 375 IU, according to daily clinical controls and estradiol values. After 12 to 16 days, follicle-stimulating hormone treatment was suspended. Within 36 to 48 hours each patient received 5000 or 10,000 IU of human chorionic gonadotropin, rarely more. Ovulation occurred in 39 of 43 treatment cycles and hyperstimulation in nine. Seven patients had normal pregnancies with viable fetuses, including one pair of twins. Two had abortions. Analysis of the endocrine situation during therapy does not permit either pregnancy or hyperstimulation to be predicted. However, hyperstimulation is frequently accompanied by endogenous luteinizing hormone peaks and greater estradiol increases during the final phase of induction. Purified follicle-stimulating hormone has thus demonstrated its validity in inducing ovulation in patients with polycystic ovarian syndrome, apparently with equal or lower risks of hyperstimulation than with other gonadotropin preparations.

The distribution and morphological behaviour of bombesin like immunoreactive cells in the proventriculus of chick embryos, newborns and adults were investigated by indirect immunoperoxidase procedure. The first immunoreactive cells appear around the 11th day of incubation. Initially few, they increase progressively to reach a peak around the 18th day. Throughout the embryonic period almost all immunoreactive cells are column-shaped, display a typical "open" behaviour, and are situated among other superficial lining epithelial cells. Around hatching time, these "open" cells suddenly decrease in number while another immunoreactive "closed" cell type becomes more evident in a deeper location. In newborns and adults, the majority of immunoreactive cells are oval shaped and are characteristically situated near the neck of lobular glands; they seem to display a "closed" behaviour. The results demonstrate two different kinds of peptide-containing cells which probably exert different functions. During the embryonic period, in fact, they resemble a chemioreceptor, but in postnatal life they appear as a paracrine/endocrine elements.

We have studied the acute effect of hOG and testosterone administration to immature rats, on testicular (particulate fraction and cytosol) and epididymal (cytosol) ABP levels. One and four hours after a single dose of 10 IU hCG/rat, ABP concentration decreased slightly in testicular membranes and cytosol, but increased markedly in epididymal cytosol (169 +/- 5 and 182 +/- 5 at 1 and 4 h respectively, compared to 113 +/- 13 fmoles/mg protein in control animals, mean +/- SEM). Since testicular and epididymal concentrations of testosterone increased after hCG, as expected, the effect might have been mediated by gonadotrophin stimulated testosterone production in Leydig cells. To test this hypothesis, the hCG effect was studied in rats that previously had received aminoglutethimide to block steroidogenesis. Under these conditions, hCG failed to increase epididymal ABP. Finally, administration of testosterone propionate was also able to stimulate epididymal ABP, but with delayed response: the increase was observed 4 h after injection (212 +/- 18 compared to 127 +/- 28 fmoles/mg protein in control animals, mean +/- SEM). It is concluded that the hCG-induced increase of intratesticular androgen levels results in rapid passage of ABP from testis to epididymis.

Testicular function was evaluated in forty-one prepubertal patients with male pseudohermaphroditism by determining serum concentrations of progesterone, 17-hydroxyprogesterone, dehydroepiandrosterone, androstenedione, testosterone and dihydrotestosterone before and after stimulation with hCG and, in some instances, ACTH. Testosterone response to hCG was normal in all subjects. In one patient, a 4-year-old boy, a deficiency of 17,20-desmolase activity was diagnosed based on the coexistence of elevated levels of pregnenolone, 17-hydroxypregnenolone, progesterone and 17-hydroxyprogesterone and low levels of dehydroepiandrosterone and androstenedione. In three other patients enzymatic blocks were suspected but not confirmed. Congenital deficiency of enzymes necessary for testosterone biosynthesis is an uncommon aetiology of male pseudohermaphroditism.

Experiments on binding 125I-LH and 125I-HCG by the human luteal tissue of early pregnancy showed no certain presence of receptors. On the other hand, studies performed with cyclic luteal tissue demonstrated the presence of high specificity, significant affinity receptors. Possible interpretations with reference to the physiological increase in HCG and/or to the characteristics of extrauterine pregnancy are discussed.

Cervical mucus forms channels when dried under a coverslip. The aim of the present work was: 1) to prove mucus canalization both in spontaneous ovulatory cycles and during ovulation induction with gonadotropins; 2) to prove the estrogen dependence of this phenomenon; 3) to check the importance of the proteidic and electrolytic concentration on chaneling; and 4) to use this phenomenon clinically, shortening the time in which it occurs. The number and arrangement of channels vary during the cycle. The phenomenon is estrogen-dependent. The comparison between estradiol values and the number of channels during spontaneous ovulatory cycles and treatment with gonadotropins showed a linear relationship. Treatment with estradiol 17 beta-valerate and ethinyl estradiol induced channel formation in women with primary amenorrhea. Canalization and ferning disappeared after dialysis or treatment with proteolytic enzymes. It follows that the two phenomena have similar characteristics. Canalization increases daily, as does estradiol, whereas ferning maintains the same grade for a longer period, and when a grade of + + + is reached, it provides no further indications. With the use of a thermostat, canalization occurred in only a few hours. Chaneling, a more precise index, could therefore substitute for ferning, particularly when monitoring the induction of ovulation.

Tissue testosterone, dihydrotestosterone, and zinc concentrations have been determined in testis and epididymis of 13 patients with carcinoma of the prostate, 1 patient with carcinoma of the penis, and 3 patients with carcinoma of the prostate on estrogens. The 13 patients not receiving estrogens had the following testicular levels: testosterone, of 529 +/- 63.1 ng/g (mean +/- SE); dihydrotestosterone, 23.7 +/- 2.58 ng/g; and zinc 62.2 +/- 7.6 micrograms/g. The epididymal levels were as follows: testosterone, 40.6 +/- 3.4 ng/g; dihydrotestosterone, 20.5 +/- 2.36 ng/g; and zinc, 67.2 +/- 11.1 micrograms/g. Patients on estrogen therapy showed much lower androgen values in the two organs but zinc was not changed significantly. Concentrations of androgens were not significantly different in the epididymal fractions of caput, corpus, and cauda. In testis, there was a positive correlation between zinc and androgens contents, while the opposite was suggested by the data in the epididymis. Even though these patients were not normal, there was no evidence of testicular or epididymal disturbances.

This study was performed on 8 patients with normal levels of PRL and ovulatory cycles. Three of these patients were treated with 5 mg/day Bromocriptine and three of them with 60 mg/day Metoclopramide from the beginning of the cycle in order to induce hypo- and hyperprolactinemia respectively. The other two women were left as control. Daily blood samples were collected for the determination of PRL, LH 17 beta-Estradiol and Progesterone. Patients were operated on between the 6th and the 8th day after the LH peak and corpora lutea enucleated and frozen for subsequent 125I-LH binding tests. An analysis of our data demonstrates that hypo- and hyper- prolactinemia induced with Bromocriptine and Metoclopramide respectively may cause luteal insufficiency, which is revealed by decreased blood levels of progesterone. Furthermore we have observed that specific binding for LH in human luteal tissue is always present (in normal, hypo- and hyperprolactinemic women) but is reduced in presence of very high or very low levels of PRL.

Androgen binding activity (ABP) was determined in two different fractions of developing rat testicular homogenates: in cytosol (cABP) and in a particulate fraction isolated by differential centrifugation (pABP). Homogenates were prepared under stabilization conditions by adding 350 nM testosterone to the homogenization buffer. cABP and pABP concentrations were maximal in 22- to 32-day-old animals, to decrease thereafter during sexual maturation. However, both cABP and pABP increased with age when results were expressed on a per organ basis. pABP could be solubilized under conditions in which it could retain its binding activity. It was then photoaffinity labeled and chromatographed on a Sephadex G 200 column using cytosolic epididymal ABP as a control. Similarities between cABP and pABP include not only the same androgen binding characteristics but also the same exclusion volume after Sephadex G 200 chromatography. Since pABP is only present in Sertoli cells, it might represent ABP before being secreted. Because of its intracellular localization, it could play a role in the compartmentalization of androgens within the testis.

Pulsatile administration of GnRH appears to be the most rational and physiological treatment of infertility in patients affected by hypothalamic amenorrhea. The authors conclude that the results obtained with this method of treatment (all patients became pregnant) suggest that the choice of pulsatile GnRH therapy is an effective and practical method for induction of ovulation.

Androgen binding activity was evaluated in different subcellular particulate fractions obtained by differential centrifugation of 32-day-old rat seminiferous tubules homogenates. After eliminating heavy particles by centrifugation at 4300 g during 4 min in 0.25 M sucrose buffer, a 27,000 g pellet was obtained and layered on 1.05 M sucrose buffer. The relatively light particulate interface (LPF) formed during centrifugation at 27,000 g 60 min, showed the highest androgen binding activity among particulate fractions. This binding was associated with the plasma membrane marker 5'-nucleotidase and it did not follow any of six other subcellular structure markers: DNA for nuclei, succinate dehydrogenase for mitochondria, acid phosphatase for lysosomes, NADPH-cytochrome C reductase for smooth endoplasmic reticulum, RNA for rough endoplasmic reticulum and lactate dehydrogenase for cytosol. In LPF, concentrations of sites were estimated to be 328 +/- 54 fmol per mg proteins and affinity constant 0.78 +/- 0.23 10(9) M-1. Heat stability, steroid specificity, affinity constant and rate of dissociation were similar to the well known androgen binding protein, ABP. Presence of ABP or a similar protein in subcellular particles might play a role in the mechanism of action of androgens in seminiferous tubules of maturing rats.

Different kidney diseases are often associated with high urinary excretion of N-acetyl-beta-D-glucosaminidase (NAG), a lysosomal enzyme involved in the breakdown of glycoproteins, whose activity is also increased in diabetic patients with poor metabolic control or vascular complications. In order to evaluate the relationship between renal function and urinary NAG levels in diabetes mellitus, 30 type II diabetic patients without evidence of kidney disease and 18 control subjects were studied. In each subject 24-h urinary excretion rates of NAG (fluorimetric method), albumin and beta 2-microglobulin (radioimmunoassay), together with 51Cr-EDTA clearance were performed. In diabetic patients urinary levels of NAG (356 +/- 25 vs 162 +/- 9.2 nmol/h/mg creatinine, p less than 0.0001) and albumin (21 +/- 2.5 vs 4.3 +/- 0.5 mg/24h, p less than 0.0001) were significantly higher than in the controls, while beta 2-microglobulin levels and 51Cr-EDTA clearance did not differ in the two groups. Moreover in diabetic patients NAG and albumin levels were positively and significantly correlated (r = 0.63, p less than 0.001). These results suggest that urinary NAG excretion rate may be altered early in diabetic patients with apparently normal renal function; its diagnostic value seems to be similar to that of the albumin excretion rate.

An androgen binding activity with characteristics similar to ABP is present in a particulate fraction (105,000 g pellet) obtained by differential centrifugation of seminiferous tubules, impure Sertoli cells and impure germ cells homogenates. Purification of germ cells through albumin gradients, results in almost a complete loss of androgen binding activity in the purified germ cell suspensions. Furthermore, Sertoli cell enriched testes from 22-, 32- and 40-day old rats showed increases in particulate androgen binding, when compared to matched controls, parallel to increments in the activity of a Sertoli cell marker enzyme (beta-glucuronidase). These results suggest that particulate androgen binding activity is only present in Sertoli cells and this protein may play a role in the compartmentalization of androgens in the testis.

The authors have incubated, in vitro, estrous rabbit endometrium with 3H-leucine and pseudopregnant rabbit endometrium with 14C-leucine. By the double-labeling technique and the use of the 14C:3H ratio, it was shown that blastokinin (BKN) is neosynthesized in vitro and that this neosynthesis probably also affects other protein components. By ion exchange chromatography of the supernatant of the pseudopregnant endometrium and previously purified BKN, it was possible to confirm the in vitro biosynthesis and to exclude interfering phenomena. Time dynamics showed that biosynthesis was more marked between the 18th and 24th hours of incubation.

Although lower esophageal sphincter (LES) dysfunction has been reported in patients with diffuse esophageal spasm, recent changes in manometric criteria for spasm and for LES relaxation suggested a need for reassessment. Moreover, LES relaxation in reflux-associated spasm has not been reported previously. On clinical criteria and independent of manometric findings, 22 patients with spasm were assigned to either idiopathic (I-DES, N = 9) or reflux-associated spasm (R-DES, N = 13) groups. Patients who underwent manometry for chest pain (C-NL, N = 10) or reflux (R-NL, N = 10) and had normal peristalsis served as control groups. Percent LES relaxation was significantly reduced in both spasm groups, and R-DES had significantly lower percent relaxation than I-DES. Post-deglutitive nadir sphincter pressure was significantly greater in R-DES than in I-DES. Duration of relaxation was normal in I-DES, but was significantly decreased in R-DES. This study indicates that 1) LES relaxation may be impaired in I-DES patients meeting current criteria for spasm, 2) the impairment in I-DES is primarily in "amplitude" of relaxation, i.e., percent relaxation and nadir pressure, but not duration, 3) LES relaxation may also be impaired in R-DES, and 4) the impairment in R-DES is to a greater degree than in I-DES patients and may be seen in both "amplitude" and duration of relaxation. This study shows that there is a spectrum of sphincter dysfunction in patients with esophageal spasm. It also suggests that there may be separate mechanisms for LES relaxation in R-DES patients, one with impaired relaxation and the other with near complete relaxation, "transient" or otherwise, to allow for reflux.

Bovine papillomavirus type 4 (BPV-4), the causative agent of alimentary papillomatosis, has been used to infect, in vitro, fragments of palatine mucosa from late term bovine fetuses. These small explants were placed beneath the renal capsule of athymic nude mice where they grew to produce, at first, squamous epithelial cysts containing BPV-4 genomic DNA and, later, papillomas which were morphologically identical to those of cattle and which contained large amounts of replicating virus. The possible utility of this technique in assessing neutralising antibodies in vaccine development is discussed.

The effect of azathioprine and inosiplex on the capacity of peripheral blood lymphocytes to form E active rosettes has been studied in vitro on 25 healthy subjects and 63 cancer patients (27 with pulmonary cancer and 36 with cancer of the digestive tract). Azathioprine significantly inhibited the capacity of T cells obtained from normal donors to form rosettes (P less than 0.01). Similar results were registered in patients suffering from pulmonary cancer (P less than 0.01) or from digestive neoplasia (P less than 0.05). Inosiplex, a substance with antiviral and immunomodulating properties, increased the percentage of the E active rosettes forming cells of 18 subjects with lung carcinoma and 26 patients with cancer of the gastrointestinal apparatus. If lymphocytes of both normal donors and cancer patients were preincubated simultaneously with azathioprine and inosiplex, the latter had an inhibitory effect on the azathioprine dependent rosette inhibition and restored the percentage of the E active rosette forming cells to control values.

Testicular testosterone concentration serum testosterone, LH and FSH, sperm count and testicular histology were evaluated in 17 patients with varicocele. Testicular testosterone was either normal or high (mean 906 +/- 723 ng/g of tissue), and serum testosterone was within the normal range in most patients. Serum LH was elevated in half of the patients. The degree of testicular damage observed was extremely variable and correlated with sperm analysis. Testicular testosterone tended to be higher in patients with severe microscopic lesions of the testis. It is concluded that even though Leydig cell function is partially altered, this deficiency is compensated by LH stimulation and therefore, failure of spermatogenesis is not secondary to low testosterone levels.

The ability of human luteal tissue to bind HLH was studied. The tissue was obtained from patients subjected to gynecological operations intended to correct other than ovarian lesions. HLH iodinated with carrier-free 125I according to the Greenwood method was used. This material has a relatively low specific radioactivity (7-20 microCi/micrograms); therefore the biological characteristics of the hormone were expected to remain intact. The 90% binding point was estimated at 37 degrees C after 5 minutes. Inhibition experiments with HLH and HCG had a characteristic effect, even those with FSH and ovine prolactin indicated a significant interference with the bound hormone. Scatchard plots demonstrated large variability in the number of binding sites and relatively modest variability of constant (Kd).

The authors studied the binding of steroids with Blastokinin (BKN). They confirm that progesterone is the steroid that has a greater affinity for BKN than estradiol or testosterone; the latter is hardly bound to the protein at all. They also show that the presence of estradiol interferes with the formation phase of the BKN-progesterone complex, while testosterone does not. When the BKN-progesterone complex is already formed, a release of the progesterone occurs only when there is a double concentration of estradiol. The biological importance of this phenomenon is briefly discussed.